Prophylactic and therapeutic treatment of infectious and other diseases with mono- and disaccharide-based compounds

ABSTRACT

Methods and compositions for treating or ameliorating diseases and other conditions, such as infectious diseases, autoimmune diseases and allergies are provided. The methods employ mono- and disaccharide-based compounds for selectively stimulating immune responses in animals and plants.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.10/757,233, filed Jan. 13, 2004, which is a continuation of U.S. patentapplication Ser. No. 09/991,376, filed Nov. 20, 2001, which is acontinuation-in-part of U.S. patent application Ser. No. 09/861,466,filed May 18, 2001, which application claims the benefit of U.S.Provisional Patent Application No. 60/281,567, filed Apr. 4, 2001, whichapplication claims the benefit of U.S. Provisional Patent ApplicationNo. 60/205,820, filed May 19, 2000, the disclosures of which areincorporated herein by reference in their entirety.

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

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BACKGROUND OF THE INVENTION

The innate immune system coordinates the inflammatory response topathogens by a system that discriminates between self and non-self viareceptors that identify classes of molecules synthesized exclusively bymicrobes. These classes are sometimes referred to as pathogen associatedmolecular patterns (PAMPs) and include, for example, lipopolysaccharide(LPS), peptidoglycans, lipotechoic acids, and bacterial lipoproteins(BLPs).

LPS is an abundant outer cell-wall constituent from gram-negativebacteria that is recognized by the innate immune system. Although thechemical structure of LPS has been known for some time, the molecularbasis of recognition of LPS by serum proteins and/or cells has onlyrecently begun to be elucidated. In a series of recent reports, a familyof receptors, referred to as Toll-like receptors (TLRs), have beenlinked to the potent innate immune response to LPS and other microbialcomponents. All members of the TLR family are membrane proteins having asingle transmembrane domain. The cytoplasmic domains are approximately200 amino acids and share similarity with the cytoplasmic domain of theIL-1 receptor. The extracellular domains of the Toll family of proteinsare relatively large (about 550-980 amino acids) and may containmultiple ligand-binding sites.

The importance of TLRs in the immune response to LPS has beenspecifically demonstrated for at least two Toll-like receptors, Tlr2 andTlr4. For example, transfection studies with embryonic kidney cellsrevealed that human Tlr2 was sufficient to confer responsiveness to LPS(Yang et al., Nature 395:284-288 (1998); Kirschning et al. J Exp Med11:2091-97 (1998)). A strong response by LPS appeared to require boththe LPS-binding protein (LBP) and CD14, which binds LPS with highaffinity. Direct binding of LPS to Tlr2 was observed at a relatively lowaffinity, suggesting that accessory proteins may facilitate bindingand/or activation of Tlr2 by LPS in vivo.

The importance of Tlr4 in the immune response to LPS was demonstrated inconjunction with positional cloning in lps mutant mouse strains. Twomutant alleles of the mouse lps gene have been identified, asemidominant allele that arose in the C3H/HeJ strain and a second,recessive allele that is present in the C57BL/10ScN and C57BL/10ScCrstrains. Mice that are homozygous for mutant alleles of lps aresensitive to infection by Gram-negative bacteria and are resistant toLPS-induced septic shock. The lps locus from these strains was clonedand it was demonstrated that the mutations altered the mouse Tlr4 genein both instances (Portorak et al., Science 282:2085-2088 (1998);Qureshi et al., J Exp Med 4:615-625 (1999)). It was concluded from thesereports that Tlr4 was required for a response to LPS.

The biologically active endotoxic sub-structural moiety of LPS islipid-A, a phosphorylated, multiply fatty-acid-acylated glucosaminedisaccharide that serves to anchor the entire structure in the outermembrane of Gram-negative bacteria. We previously reported that thetoxic effects of lipid A could be ameliorated by selective chemicalmodification of lipid A to produce monophosphoryl lipid A compounds(MPL® immunostimulant; Corixa Corporation; Seattle, Wash.). Methods ofmaking and using MPL® immunostimulant and structurally like compounds invaccine adjuvant and other applications have been described (see, forexample, U.S. Pat. Nos. 4,436,727; 4,877,611; 4,866,034 and 4,912,094;4,987,237; Johnson et al., J Med Chem 42:4640-4649 (1999); Ulrich andMyers, in Vaccine Design: The Subunit and Adjuvant Approach; Powell andNewman, Eds.; Plenum: New York, 495-524, 1995; the disclosures of whichare incorporated herein by reference in their entireties). Inparticular, these and other references demonstrated that MPL®immunostimulant and related compounds had significant adjuvantactivities when used in vaccine formulations with protein andcarbohydrate antigens for enhancing humoral and/or cell-mediatedimmunity to the antigens.

Moreover, we have previously described a class of synthetic mono- anddisaccharide mimetics of monophosphoryl lipid A, referred to asaminoalkyl glucosaminide phosphates (AGPs), for example in U.S. Ser. No.08/853,826, now U.S. Pat. No. 6,113,918, Ser. Nos. 09/074,720,09/439,839, now U.S. Pat. No. 6,303,347, and in PCT/US98/09385 (WO98/50399, Oct. 12, 1998) the disclosures of which are incorporatedherein by reference in their entireties. Like monophosphoryl lipid A,these compounds have been demonstrated to retain significant adjuvantcharacteristics when formulated with antigens in vaccine compositionsand, in addition, have similar or improved toxicity profiles whencompared with monophosphoryl lipid A. A significant advantage offered bythe AGPs is that they are readily producible on a commercial scale bysynthetic means.

Although monophosphoryl lipid A and the AGPs have been describedprimarily for use in combination with antigens in vaccine formulations,their use as monotherapies, in the absence of antigen, for theprophylactic and/or therapeutic treatment of plant and animal diseasesand conditions, such as infectious disease, autoimmunity and allergies,has not been previously reported.

The present invention, as a result of a growing understanding of certainmechanisms underlying the activities of monophosphoryl lipid A and AGPcompounds, makes possible the novel therapeutic opportunities describedherein.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention provides methods for treating,ameliorating or substantially preventing a disease or condition in ananimal by administering an effective amount of a compound having theformula:

and pharmaceutically acceptable salts thereof, wherein X is —O— or —NH—;R¹ and R² are each independently a (C₂-C₂₄)acyl group, includingsaturated, unsaturated and branched acyl groups; R³ is —H or —PO₃R¹¹R¹²,wherein R¹¹ and R¹² are each independently-H or (C₁-C₄)alkyl; R⁴ is —H,—CH₃ or —PO₃R¹³R¹⁴, wherein R¹³ and R¹⁴ are each independently selectedfrom —H and (C₁-C₄)alkyl; and Y is a radical selected from the formulae:

wherein the subscripts n, m, p and q are each independently an integerof from 0 to 6; R⁵ is a (C₂-C₂₄)acyl group (including, as above,saturated, unsaturated and branched acyl groups); R⁶ and R⁷ areindependently selected from H and CH₃; R⁸ and R⁹ are independentlyselected from H, OH, (C₁-C₄)alkoxy, —PO₃H₂, —OPO₃H₂, —SO₃H, —OSO₃H,—NR¹⁵R¹⁶, —SR¹⁵, —CN, —NO₂, —CHO, —CO₂R¹⁵, —CONR¹⁵R¹⁶, —PO₃R¹⁵R¹⁶,—OPO₃R¹⁵R¹⁶, —SO₃R¹⁵ and —OSO₃R¹⁵, wherein R¹⁵ and R¹⁶ are eachindependently selected from H and (C₁-C₄)alkyl; R¹⁰ is selected from H,CH₃, —PO₃H₂, ω-phosphonooxy(C₂-C₂₄)alkyl, and ω-carboxy(C₁-C₂₄)alkyl;and Z is —O— or —S—; with the proviso that when R³ is —PO₃R¹¹R¹², R⁴ isother than —PO₃R¹³R¹⁴.

In certain illustrative aspects of the invention, the above methods areemployed in treating, ameliorating or substantially preventinginfectious diseases, autoimmune diseases and allergies.

The present invention, in other aspects, provides pharmaceuticalcompositions comprising one or more of the compounds described above ina suitable excipient, formulated and/or administered in the absence ofexogenous antigen.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph depicting the nonspecific protection of mice againstlethal influenza challenge following or coincident with MonoPhosphorylLipid A (MPL) administration.

FIG. 2 is a graph depicting clinical symptoms following intranasaladministration of L-Seryl Aminoalkyl Glucosaminide Phosphates (AGPs) tomice.

FIG. 3 is a graph depicting clinical symptoms following L-SerylAminoalkyl Glucosaminide Phosphates (AGPs) monotherapy and influenzachallenge.

FIGS. 4-6 are graphs depicting cytokine induction by RC522 as comparedto MPL in overnight whole blood cultures from three human donors (donorsA-C, respectively).

FIG. 7 are graphs depicting cytokine induction by RC522 as compared toMPL in short term whole blood cultures from donor A.

FIG. 8 are graphs depicting cytokine induction by RC522 as compared toMPL in murine (Balb/c and C3H/HEJ) splenic cultures.

FIG. 9 are graphs depicting cytokine induction by RC529 and RC552 ascompared to MPL in human peripheral blood mononuclear cells (PBMC).

FIG. 10 A-E is a figure showing various AGP compounds.

FIG. 11 is a graph showing clinical symptoms following intranasaladministration of L-Seryl Aminoalkyl Glucosaminide Phosphatesmonotherapy and influenza challenge.

FIG. 12 is a graph showing clinical symptoms following intranasaladministration of L-Seryl 666 versus L-Seryl 000 AminoalkylGlucosaminide Phosphates monotherapy and influenza challenge.

FIG. 13 is a graph showing clinical symptoms following intranasaladministration of L-Seryl Aminoalkyl Glucosaminide Phosphatesmonotherapy and influenza challenge. L-Seryl compounds have variouscombinations of 6 and 10 carbon fatty acid chains in the secondary fattyacid position.

FIG. 14 is a graph showing clinical symptoms following intravenousadministration of L-Seryl Aminoalkyl Glucosaminide Phosphatesmonotherapy and intravenous Listeria monocytogenes challenge.

FIG. 15 is a graph showing clinical symptoms following intravenousadministration of L-Seryl Aminoalkyl Glucosaminide Phosphatesmonotherapy and intravenous Listeria monocytogenes challenge. L-Serylcompounds have various combinations of 6 and 10 carbon fatty acid chainsin the secondary fatty acid position.

FIG. 16 is a graph showing clinical symptoms following intravenousadministration of various Aminoalkyl Glucosaminide Phosphatesmonotherapy and intravenous Listeria monocytogenes challenge. The AGPcompounds all have 10 carbon fatty acid chains in the secondary fattyacid position.

FIG. 17 is a graph showing clinical symptoms following intravenousadministration of Aminoalkyl Glucosaminide Phosphates monotherapy andintravenous Listeria monocytogenes challenge. The AGP compounds vary inlinker length between the glucosamine and aglycone moieties.

DETAILED DESCRIPTION OF THE INVENTION

Illustrative Prophylactic and Therapeutic Applications

The present invention broadly concerns prophylactic and therapeuticmethods of treating certain diseases and other medical conditions byadministration of an effective amount of one or more mono- ordisaccharide compounds described herein or a pharmaceutical compositioncomprising one or more such compounds. While certain of the mono- anddisaccharide compounds have been described for use as adjuvants incombination with exogenously administered antigens in vaccineformulations, and for use in certain other applications, the presentinvention provides novel therapeutic methods that employ the compoundspreferably in monotherapeutic applications, i.e., in the absence ofexogenously administered antigen.

Thus, in one aspect, the present invention provides methods fortreating, ameliorating and/or substantially preventing infectiousdiseases in eukaryotic subjects, particularly in animals, preferably inhumans. Given the importance of TLR-mediated signalling in the innateimmune response to microbial challenge, the ability to stimulate suchpathways selectively and with minimal toxicity represents a powerfulapproach for prophylactic and/or therapeutic treatment modalitiesagainst a wide range of infectious agents.

The methods described herein are applicable against essentially any typeof infectious agent, including bacteria, viruses, parasites, and fungi.Illustratively, the invention is useful for the prophylactic and/ortherapeutic treatment of bacterial infections by species fromPseudomonas, Escherichia, Klebsiella, Enterobacter, Proteus, Serratia,Candida, Staphylococci, Streptococci, Chlamydia, Mycoplasma and numerousothers. Illustrative viral conditions that may be treated in accordancewith the invention include those caused, for example, by Influenzaviruses, Adenoviruses, parainfluenza viruses, Rhinoviruses, respiratorysyncytial viruses (RSVs), Herpes viruses, Cytomegaloviruses, Hepatitisviruses, e.g., Hepatitis B and C viruses, and others. Illustrative fungiinclude, for example, Aspergillis, Candida albicans, Cryptococcusneoformans, Coccidioides immitus, and others.

In one illustrative embodiment, the invention provides methods for thetreatment of subjects, particularly immunocompromised subjects, thathave developed or are at risk for developing infections, such asnosocomial bacterial and viral infections. About 2 million of the 40million individuals hospitalized every year develop nosocomial infectionduring their stay and about 1% of these, or about 400,000 patients,develop nosocomial pneumonia, more than 7000 of which die. This makesnosocomial pneumonia the leading cause of death in hospital-acquiredinfections. Thus, this embodiment fills a significant need for effectiveprophylactic approaches in the treatment of nosocomial infections.

In a related embodiment, the present invention provides prophylactictreatments for immunocompromised patients, such as HIV-positivepatients, who have developed or are at risk for developing pneumoniafrom either an opportunistic infection or from the reactivation of asuppressed or latent infection. In 1992, about 20,000 cases ofPneumocystis carinii infections in AIDS patients were reported in theU.S. alone. Additionally, 60-70% of all AIDS patients get P. carinii atsome time during their illness. Thus, the present invention in thisembodiment provides effective prophylactic methods for this at-riskpopulation.

In another related embodiment, the methods of the present invention areused for treating other patient populations that may beimmunocompromised and/or at risk for developing infectious diseases,including, for example, patients with cystic fibrosis, chronicobstructive pulmonary disease and other immunocompromized and/orinstitutionalized patients.

In support of these and other embodiments of the invention, we havedemonstrated that pre-challenge administration of an illustrativecompound of the present invention in immunocompromised mice providessignificant prophylactic protection against infection by Pneumocystiscarinii. (See Example 1).

In another aspect of the invention, the mono- and disaccharide compoundsdescribed herein are employed in methods for treating, ameliorating orsubstantially preventing allergic disorders and conditions, such assinusitis, chronic rhinosinusitus, asthma, atopic dermatitis andpsoriasis. This approach is based at least in part on the ability of themono- and disaccharide compounds to activate the production of cytokinesfrom target cells that can compete with stereotypic allergic-typecytokine responses characterized by IL-4 production orhyperresponsiveness to IL-4 activity. Administration of certain of themono- and disaccharide compounds disclosed herein results in IFN-gammaand IL-12 expression from antigen processing and presenting cells, aswell as other cells, resulting in down regulation of cytokinesassociated with allergic responses such as IL-4, 5, 6, 10 and 13.

In another aspect of the invention, mono- and disaccharide compounds areemployed in methods for treating autoimmune diseases and conditions. Themono- and disaccharide compounds for use in this embodiment willtypically be selected from those capable of antagonizing, inhibiting orotherwise negatively modulating one or more Toll-like receptors,particularly Tlr2 and/or Tlr4, such that an autoimmune responseassociated with a given condition is ameliorated or substantiallyprevented. Illustratively, the methods provided by this embodiment canbe used in the treatment of conditions such as inflammatory boweldisease, rheumatoid arthritis, chronic arthritis, multiple sclerosis andpsoriasis.

While not wishing to be bound by theory, it is believed that theefficacy of the prophylactic and therapeutic applications describedabove are based at least in part on the involvement of the mono- anddisaccharide compounds in the modulation of Toll-like receptor activity.In particular, Toll-like receptors Tlr2, Tlr4, and others, are believedto be specifically activated, competitively inhibited or otherwiseaffected by the non-toxic LPS derivatives and mimetics disclosed herein.Accordingly, the methods of the invention provide a powerful andselective approach for modulating the innate immune response pathways inanimals without giving rise to the toxicities often associated with thenative bacterial components that normally stimulate those pathways.

Illustrative Mono- and Disaccharide Compounds

Illustrative mono- or disaccharide compounds employed in the aboveprophylactic and therapeutic applications comprise compounds having theformula:

and pharmaceutically acceptable salts thereof, wherein X is —O— or —NH—;R¹ and R² are each independently a (C₂-C₂₄)acyl group, includingsaturated, unsaturated and branched acyl groups; R³ is —H or —PO₃R¹¹R¹²,wherein R¹¹ and R¹² are each independently-H or (C₁ C₄)alkyl; R⁴ is —H,═CH₃ or —PO3R¹³R¹⁴, wherein R¹³ and R¹⁴ are each independently selectedfrom —H and (C₁-C₄)alkyl; and Y is a radical selected from the formulae:

wherein the subscripts n, m, p and q are each independently an integerof from 0 to 6; R⁵ is a (C₂-C₂₄)acyl group (including, as above,saturated, unsaturated and branched acyl groups); R⁶ and R⁷ areindependently selected from H and CH₃; R⁸ and R⁹ are independentlyselected from H, OH, (C₁-C₄)alkoxy, —PO₃H₂, —OPO₃H₂, —SO₃H, —OSO₃H,—NR¹⁵R¹⁶, —SR⁵, —CN, —NO₂, —CHO, —CO₂R⁵, —CONR¹⁵R¹⁶, PO₃R¹⁵R¹⁶,—OPO₃R¹⁵R¹⁶, —SO₃R¹⁵ and —OSO₃R¹⁵, wherein R¹⁵ and R¹⁶ are eachindependently selected from H and (C₁-C₄)alkyl; R¹⁰ is selected from H,CH₃, —PO₃H₂, ω-phosphonooxy(C₂-C₂₄)alkyl, and ω-carboxy(C₁-C₂₄)alkyl;and Z is —O— or —S; with the proviso that when R³ is —PO₃R¹¹R¹², R⁴ isother than —PO₃R³R⁴.

Additionally, when R³ is —PO₃H₂, R⁴ is H, R¹⁰ is H, R¹ isn-tetradecanoyl, R² is n-octadecanoyl and R⁵ is n-hexadecanoyl, then Xis other than —O.

In the general formula above, the configuration of the 3′ stereogeniccenters to which the normal fatty acid acyl residues are attached is Ror S, but preferably R. The stereochemistry of the carbon atoms to whichR⁶ and R⁷ are attached can be R or S. All stereoisomers, enantiomers,diastereomers and mixtures thereof are considered to be within the scopeof the present invention.

In one group of preferred embodiments, Y has the formula:

Within this group of embodiments, the acyl groups R¹, R² and R⁵ will beselected such that at least two of the groups are (C₂-C₆)acyl. Furtherpreferred are those embodiments in which the total number of carbonatoms in R¹, R² and R⁵ is from about 6 to about 22, more preferablyabout from about 12 to about 18. In other preferred embodiments, X is Oand Z is O. The subscripts n, m, p and q are preferably integers of from0 to 3, more preferably, 0 to 2. Of the remaining substituents, R⁶ andR⁷ are preferably H. The present invention further contemplates thoseembodiments in which the preferred substituents are combined in onemolecule.

In another group of embodiments, R¹, R² and R⁵ are selected from(C₁₂-C₂₀)acyl with the proviso that the total number of carbon atoms inR¹, R² and R⁵ is from about 44 to about 60. More preferably, the totalnumber of carbon atoms in R¹, R² and R⁵ is from about 46 to about 52.Still further preferred are those embodiments in which X and Z are both—O—.

In another group of embodiments, Y has the formula:

As with the preferred group of embodiments provided above, in this groupthe acyl groups R¹, R² and R⁵ will also be selected such that at leasttwo of the groups are (C₂-C₆)acyl. Further preferred are thoseembodiments in which the total number of carbon atoms in R¹, R² and R⁵is from about 6 to about 22, more preferably about from about 12 toabout 18. In other preferred embodiments, X is O. Of the remainingsubstituents, R³ is preferably phosphono (—PO₃H₂) and R⁴ is preferablyH. The present invention further contemplates those embodiments in whichvarious combinations of the preferred substituents are combined in onemolecule.

In another group of embodiments, R¹, R² and R⁵ are selected from(C₁₂-C₂₄)acyl with the proviso that the total number of carbon atoms inR¹, R² and R⁵ is from about 44 to about 60. More preferably, the totalnumber of carbon atoms in R¹, R² and R⁵ is from about 46 to about 52.Particularly preferred fatty acid groups for R¹, R² and R⁵ are normalC₁₄, C₁₆ and C₁₈ fatty acid groups. Still further preferred are thoseembodiments in which X is —O—. Similar to the shorter acyl chainembodiments provided above, R³ is preferably phosphono (—PO₃H₂) and R⁴is preferably H.

In another preferred embodiments of the present invention, Y is aradical of formula (Ib), X is O, R³ is phosphono, R⁴ is H, and R¹, R²and R⁵ are selected from (C₁₂-C₂₄)acyl with the proviso that the totalnumber of carbon atoms in R¹, R² and R⁵ is from about 46 to about 52.Still further preferred are those compounds in which R² is(C₁₆-C₁₈)acyl.

The term “alkyl” by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain, or cyclichydrocarbon radical, or combination thereof, which may be fullysaturated, mono- or polyunsaturated and can include di- and multivalentradicals, having the number of carbon atoms designated (i.e., C₁-C₁₀means one to ten carbons). Examples of saturated hydrocarbon radicalsinclude groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl,t-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl,cyclopropylmethyl, homologs and isomers of, for example, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. An unsaturated alkyl group isone having one or more double bonds or triple bonds. Examples ofunsaturated alkyl groups include vinyl, 2-propenyl, crotyl,2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl),ethynyl, 1- and 3-propynyl, 3-butynyl, and the higher homologs andisomers. Typically, an alkyl group will have from 1 to 24 carbon atoms.A “lower alkyl” or is a shorter chain alkyl group, generally havingeight or fewer carbon atoms.

The terms “alkoxy”, “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively.

The term “acyl” refers to a group derived from an organic acid byremoval of the hydroxy group. Examples of acyl groups include acetyl,propionyl, dodecanoyl, tetradecanoyl, isobutyryl, and the like.Accordingly, the term “acyl” is meant to include a group otherwisedefined as —C(O)-alkyl.

Each of the above terms (e.g., “alkyl” “acyl”) are meant to include bothsubstituted and unsubstituted forms of the indicated radical. Preferredsubstituents for each type of radical are provided below.

Substituents for the alkyl and acyl radicals can be a variety of groupsselected from: —OR′, ═O, ═NR′, ═N—OR′, —NR′R″, —SR′, -halogen,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NHC(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —CN and —NO₂ in a numberranging from zero to (2m′+1), where m′ is the total number of carbonatoms in such radical. R′, R″ and R′″ each independently refer tohydrogen and unsubstituted (C₁-C₈)alkyl. When R′ and R″ are attached tothe same nitrogen atom, they can be combined with the nitrogen atom toform a 5-, 6-, or 7-membered ring. For example, —NR′R″ is meant toinclude 1-pyrrolidinyl and 4-morpholinyl. From the above discussion ofsubstituents, one of skill in the art will understand that the term“alkyl” is meant to include groups such as haloalkyl (e.g., —CF₃ and—CH₂CF₃) and the like.

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, maleic, malonic, benzoic, succinic,suberic, fumaric, lactic, mandelic, phthalic, benzenesulfonic,p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like. Alsoincluded are salts of amino acids such as arginate and the like, andsalts of organic acids like glucuronic or galactunoric acids and thelike (see, for example, Berge, S. M., et al, “Pharmaceutical Salts,”Journal of Pharmaceutical Science, 66, 1-19, 1977). Certain specificcompounds of the present invention contain both basic and acidicfunctionalities that allow the compounds to be converted into eitherbase or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers and individual isomers are all intended to beencompassed within the scope of the present invention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (125I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

The mono- and disaccharide compounds can be prepared by any suitablemeans, many of which have been described. For example, certain compoundsuseful in the present invention are described in application Ser. Nos.08/853,826, now U.S. Pat. No. 6,113,918; Ser. No. 09/439,839 (filed Nov.12, 1999), now U.S. Pat. No. 6,303,347; and in PCT/US98/09385 (WO98/50300, Oct. 12, 1998) the disclosures of which are incorporatedherein by reference in their entireties. Other compounds can be preparedin a manner similar to that described for RC-552 (L34) in U.S. Pat. No.6,013,640. Still other compounds can be prepared using methods outlinedin Johnson, et al., J. Med. Chem. 42:4640-4649 (1999), Johnson, et al.,Bioorg. Med. Chem. Lett. 9:2273-2278 (1999), and PCT/US98/50399 (WO98/50399, Nov. 12, 1998). Still other compounds can be preparedaccording to, for example, U.S. Pat. Nos. 4,436,727; 4,877,611;4,866,034; 4,912,094; and 4,987,237. In general, the synthetic methodsdescribed in the above-noted references and other synthetic methodsotherwise familiar in the art are broadly applicable to the preparationthese compounds. For example, in making compounds having different acylgroups and substitutions, one of skill in the art will appreciate thatthe convergent methods described therein can be modified to usealternate acylating agents, or can be initiated with commerciallyavailable materials having appropriate acyl groups attached.

Illustrative Pharmaceutical Compositions and their Delivery

In another embodiment, the present invention concerns pharmaceuticalcompositions comprising one or more of the mono- and disaccharidecompounds disclosed herein in pharmaceutically-acceptablecarriers/excipients for administration to a cell, tissue, animal orplant, either alone, or in combination with one or more other modalitiesof therapy. In a preferred embodiment, the pharmaceutical compositionsare formulated in the absence of exogenous antigen, i.e., are used inmonotherapeutic applications. For many such embodiments, thepharmaceutical compositions of the invention will comprise one or moreof the monosaccharide compounds described herein.

Illustrative carriers for use in formulating the pharmaceuticalcompositions include, for example, oil-in-water or water-in-oilemulsions, aqueous compositions with or without inclusion of organicco-solvents suitable for intravenous (IV) use, liposomes orsurfactant-containing vesicles, microspheres, microbeads and microsomes,powders, tablets, capsules, suppositories, aqueous suspensions,aerosols, and other carriers apparent to one of ordinary skill in theart.

In certain embodiments, the pharmaceutical compositions will compriseone or more buffers (e.g., neutral buffered saline or phosphate bufferedsaline), carbohydrates (e.g., glucose, mannose, sucrose or dextrans),mannitol, proteins, polypeptides or amino acids such as glycine,antioxidants, bacteriostats, chelating agents such as EDTA orglutathione, adjuvants (e.g., aluminum hydroxide), solutes that renderthe formulation isotonic, hypotonic or weakly hypertonic with the bloodof a recipient, suspending agents, thickening agents and/orpreservatives.

For certain applications, aqueous formulations will be preferred,particularly those comprising an effective amount of one or moresurfactants. For example, the composition can be in the form of amicellar dispersion comprising at least one suitable surfactant, e.g., aphospholipid surfactant. Illustrative examples of phospholipids includediacyl phosphatidyl glycerols, such as dimyristoyl phosphatidyl glycerol(DPMG), dipalmitoyl phosphatidyl glycerol (DPPG), and distearoylphosphatidyl glycerol (DSPG), diacyl phosphatidyl cholines, such asdimyristoyl phosphatidylcholine (DPMC), dipalmitoyl phosphatidylcholine(DPPC), and distearoyl phosphatidylcholine (DSPC); diacyl phosphatidicacids, such as dimyristoyl phosphatidic acid (DPMA), dipalmitoylphosphatidic acid (DPPA), and distearoyl phosphatidic acid (DSPA); anddiacyl phosphatidyl ethanolamines such as dimyristoyl phosphatidylethanolamine (DPME), dipalmitoyl phosphatidyl ethanolamine (DPPE) anddistearoyl phosphatidyl ethanolamine (DSPE). Typically, asurfactant:mono-/disaccharide molar ratio in an aqueous formulation willbe from about 10:1 to about 1:10, more typically from about 5:1 to about1:5, however any effective amount of surfactant may be used in anaqueous formulation to best suit the specific objectives of interest.

The compounds and pharmaceutical compositions of the invention can beformulated for essentially any route of administration, e.g., injection,inhalation by oral or intranasal routes, rectal, vaginal orintratracheal instillation, ingestion, or transdermal or transmucosalroutes, and the like. In this way, the therapeutic effects attainable bythe methods and compositions of the invention can be, for example,systemic, local, tissue-specific, etc., depending of the specific needsof a given application of the invention.

Illustrative formulations can be prepared and administered parenterally,i.e., intraperitoneally, subcutaneously, intramuscularly orintravenously. One illustrative example of a carrier for intravenous useincludes a mixture of 10% USP ethanol, 40% USP propylene glycol orpolyethylene glycol 600 and the balance USP Water for Injection (WFI).Other illustrative carriers include 10% USP ethanol and USP WFI;0.01-0.1% triethanolamine in USP WFI; or 0.01-0.2% dipalmitoyldiphosphatidylcholine in USP WFI; and 1-10% squalene or parenteralvegetable oil-in-water emulsion. Pharmaceutically acceptable parenteralsolvents will generally be selected such that they provide a solution ordispersion which may be filtered through a 0.22 micron filter withoutremoving the active ingredient.

Illustrative examples of carriers for subcutaneous or intramuscular useinclude phosphate buffered saline (PBS) solution, 5% dextrose in WFI and0.01-0.1% triethanolamine in 5% dextrose or 0.9% sodium chloride in USPWFI, or a 1 to 2 or 1 to 4 mixture of 10% USP ethanol, 40% propyleneglycol and the balance an acceptable isotonic solution such as 5%dextrose or 0.9% sodium chloride; or 0.01-0.2% dipalmitoyldiphosphatidylcholine in USP WFI and 1 to 10% squalene or parenteralvegetable oil-in-water emulsions.

Examples of carriers for administration via mucosal surfaces depend uponthe particular route, e.g., oral, sublingual, intranasal, etc. Whenadministered orally, illustrative examples include pharmaceutical gradesof mannitol, starch, lactose, magnesium stearate, sodium saccharide,cellulose, magnesium carbonate and the like, with mannitol beingpreferred. When administered intranasally, illustrative examples includepolyethylene glycol, phospholipids, glycols and glycolipids, sucrose,and/or methylcellulose, powder suspensions with or without bulkingagents such as lactose and preservatives such as benzalkonium chloride,EDTA. In a particularly illustrative embodiment, the phospholipid 1,2dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) is used as an isotonicaqueous carrier at about 0.01-0.2% for intranasal administration of thecompound of the subject invention at a concentration of about 0.1 to 3.0mg/ml.

When administered by inhalation, illustrative carriers includepolyethylene glycol or glycols, DPPC, methylcellulose, powdereddispersing agents, and preservatives, with polyethylene glycols and DPPCbeing preferred. In many instances, it will be preferred that the mono-or disaccharide compounds be in a nebulized form when administration byinhalation. Illustratively, delivery may be by use of a single-usedelivery device, a mist nebulizer, a breath-activated powder inhaler, anaerosol metered-dose inhaler (MDI) or any other of the numerousnebulizer delivery devices available in the art. Additionally, misttents or direct administration through endotracheal tubes may also beused. Delivery via an intratracheal or nasopharyngeal mode will beefficacious for certain indications.

One skilled in this art will recognize that the above description isillustrative rather than exhaustive. Indeed, many additionalformulations techniques and pharmaceutically-acceptable excipients andcarrier solutions are well-known to those skilled in the art, as is thedevelopment of suitable dosing and treatment regimens for using theparticular compositions described herein in a variety of treatmentregimens.

The compounds can be evaluated in a variety of assay formats to identifyand select those having the characteristics best suited for a givenapplication of the invention. For example, animal models can be used foridentifying and evaluating cytokine release profiles into systemiccirculation following administration of a mono- and/or disaccharidecompound. In addition, various in vitro and in vivo models exist forexamining changes in one or more aspects of an immune response todifferent antigenic components in order to identify compounds bestsuited for eliciting a specific immune response of interest. Forexample, a compound can be contacted with target cells, such asmacrophages, dendritic cells or Langerhans cells in vitro, andelaborated cytokines can be measured. In addition, gene expressionarrays can be used to identify specific pathways activated or inhibitedby a particular mono- or disaccharide of interest.

It will be understood that, if desired, the compounds disclosed hereinmay be administered in combination with other therapeutic modalities,such as antimicrobial, antiviral and antifungal compounds or therapies,various DNA-based therapeutics, RNA-based therapeutics,polypeptide-based therapeutics, and/or with other immunoeffectors. Infact, essentially any other component may also be included, given thatthe additional component(s) do not cause a significant adverse effectupon contact with the target cells or host tissues. The compositions maythus be delivered along with various other agents as required or desiredfor the specific embodiment(s) of the invention being implemented.

Illustratively, the pharmaceutical compositions of the invention caninclude, or be used in conjunction with, DNA encoding one or moretherapeutic proteins, antisense RNAs, ribozymes or the like. The DNA maybe present within any of a variety of delivery systems known to those ofordinary skill in the art, including nucleic acid expression systems,bacteria and viral expression systems. Numerous gene delivery techniquesare well known in the art, such as those described by Rolland, Crit.Rev. Therap. Drug Carrier Systems 15:143-198, 1998, and references citedtherein. Appropriate nucleic acid expression systems contain thenecessary DNA sequences for expression in the patient (such as asuitable promoter and terminating signal). In a preferred embodiment,the DNA may be introduced using a viral expression system (e.g.,vaccinia or other pox virus, retrovirus, or adenovirus), which mayinvolve the use of a non-pathogenic (defective), replication competentvirus. Suitable systems are disclosed, for example, in Fisher-Hoch etal., Proc. Natl. Acad. Sci. USA 86:317-321, 1989; Flexner et al., Ann.N.Y. Acad. Sci. 569:86 103, 1989; Flexner et al., Vaccine 8:17 21, 1990;U.S. Pat. Nos. 4,603,112, 4,769,330, and 5,017,487; WO 89/01973; U.S.Pat. No. 4,777,127; GB 2,200,651; EP 0,345,242; WO 91/02805; Berkner,Biotechniques 6:616 627, 1988; Rosenfeld et al., Science 252:431-434,1991; Kolls et al., Proc. Natl. Acad. Sci. USA 91:215 219, 1994; KassEisler et al., Proc. Natl. Acad. Sci. USA 90:11498 11502, 1993; Guzmanet al., Circulation 88:2838 2848, 1993; and Guzman et al., Cir. Res.73:1202 1207, 1993. Techniques for incorporating DNA into suchexpression systems are well known to those of ordinary skill in the art.

The DNA may also be “naked,” as described, for example, in Ulmer et al.,Science 259:1745-1749, 1993 and reviewed by Cohen, Science259:1691-1692, 1993. The uptake of naked DNA may be increased by coatingthe DNA onto biodegradable beads, which are efficiently transported intothe cells. It will be apparent that a pharmaceutical composition of theinvention may comprise both a polynucleotide and a protein component.

Any of a variety of additional immunostimulants may be included in thecompositions of this invention. For example, cytokines, such as GM-CSF,interferons or interleukins to further modulate an immune response ofinterest. For example, in certain embodiments, additional components maybe included in the compositions to further enhance the induction of highlevels of Th1-type cytokines (e.g., IFN-γ, TNFα, IL-2 and IL-12).Alternatively, or in addition, high levels of Th2-type cytokines (e.g.,IL-4, IL-5, IL-6 and IL-10) may be desired for certain therapeuticapplications. The levels of these cytokines may be readily assessedusing standard assays. For a review of the families of cytokines, seeMosmann and Coffman, Ann. Rev. Immunol. 7:145-173, 1989.

Illustrative compositions for use in induction of Th1-type cytokinesinclude, for example, a combination of CpG-containing oligonucleotides(in which the CpG dinucleotide is unmethylated) as described, forexample, in WO 96/02555, WO 99/33488 and U.S. Pat. Nos. 6,008,200 and5,856,462. Immunostimulatory DNA sequences are also described, forexample, by Sato et al., Science 273:352, 1996. Other suitableimmunostimulants comprise saponins, such as QS21 (AquilaBiopharmaceuticals Inc., Framingham, Mass.), and related saponinderivatives and mimetics thereof.

Other illustrative immunostimulants that can be used in conjunction withthe present invention include Montamide ISA 720 (Seppic, France), SAF(Chiron, Calif., United States), ISCOMS (CSL), MF-59 (Chiron), the SBASseries of adjuvants (e.g., SBAS-2 or SBAS-4, available from SmithKlineBeecham, Rixensart, Belgium), and Enhanzyn™ immunostimulant (Corixa,Hamilton, Mont.). Polyoxyethylene ether immunostimulants, are describedin WO 99/52549A1.

General Definitions:

As used herein, “an effective amount” is that amount which shows aresponse over and above the vehicle or negative controls. As discussedabove, the precise dosage of the compound of the subject invention to beadministered to a patient will depend the route of administration, thepharmaceutical composition, and the patient.

The phrase “pharmaceutically acceptable” refers to molecular entitiesand compositions that do not produce an allergic or similar untowardreaction when administered to a human.

As used herein, “carrier” or “excipient” includes any and all solvents,dispersion media, vehicles, coatings, diluents, antibacterial andantifungal agents, isotonic and absorption delaying agents, buffers,carrier solutions, suspensions, colloids, and the like. The use of suchmedia and agents for pharmaceutical active substances is well known inthe art. Except insofar as any conventional media or agent isincompatible with the active ingredient, its use in the therapeuticcompositions is contemplated.

EXAMPLES Example 1 Protection Against P. carinii Infection byProphylactic Administration of MonoPhosphoryl Lipid A

Mice were pretreated with L3T4 anti-CD4 antibody for a minimum of 2weeks (2 injections/week, 0.2 mg/injection) or until the peripheral CD4count was reduced by at least about 50%.

An aqueous formulation was prepared containing 1 mg/ml 3-O-deacylatedmonophosphoryl lipid A and 108 μg/ml of the surfactant DPPC in water.The formulation was administered intratracheally via a small cannula at−24 hours and then twice a week for the remainder of the study. Theconcentrations administered are indicated below in Table 1. One millionP. carinii were inoculated trans-tracheally at day 0. Twice-weeklytreatments were continued for 7 weeks, lungs were removed and impressionsmears made. Slides were stained with Giemsa and silver and scored forthe presence of P. carinii as follows.

Score 5 >100/1000x field 4 10-100/field 3 1-10/field 2 1-10/10 fields 11-10/50 fields 0 0/50 fieldsThe results of these experiments are summarized below in Table 1:

TABLE 1 GIEMSA SILVER PLACEBO GROUP 3.3 ± 0.7 2.5 ± 0.4  25 MG/KG 3.4 ±0.5 3.3 ± 0.1 100 MG/KG 2.7 ± 0.5 3.2 ± 0.1 250 MG/KG 1.8 ± 0.8 1.6 ±0.2This study was repeated and the following results were obtained (Table2):

TABLE 2 GIEMSA PLACEBO GROUP 3.3 ± 0.2 UNTREATED CONTROL 3.1 ± 0.3 100MG/KG 1.3 ± 0.3 200 MG/KG 1.0 ± 0.3

These results demonstrate that pulmonary delivery of monophosphoryllipid A promotes nonspecific resistance to infection by Pneumocystiscarinii in immunocompromised mice. Inhalation of monophosphoryl lipid Aled to activation of the local (and distal) innate immune responsesresulting in enhanced nonspecific protection. Monophosphoryl lipid Amediated this protection primarily through activation of antigenpresenting cells leading to increased phagocytic activity and therelease of immuno-stimulatory cytokines. FACS analysis of cell lavagedfrom the lungs displayed markers for activated neutrophils but wasunremarkable for an influx of leukocytes characteristic of a massiveinflammatory response (ARDS). Analysis of spleen cells showed negativeexpression of CD11b or CD69, suggesting that the monophosphoryl lipid Aformulations and the effects in this application were not systemic butwere confined to the lung.

Example 2 Protection Against Lethal Influenza Challenge by ProphylacticAdministration of MonoPhosphoryl Lipid A

A dose of 20 μg MonoPhosphoryl Lipid A (MPL) was given to groups offemale BALB/c mice by intranasal (i.n.) administration either 2 daysprior to or the day of lethal influenza challenge. All mice werechallenged with approximately 2 LD50 infectious influenza A/HK/68administered i.n. Mortality was monitored for 21 days followinginfluenza challenge. The results of these experiments is presented inFIG. 1. These data demonstrate that intranasal delivery ofmonophosphoryl lipid A promotes nonspecific resistance to infection bylethal influenza challenge in mice.

Example 3 Clinical Symptoms Following Intranasal Administration ofL-Seryl Aminalkyl Glucosaminide Phosphates (AGPS)

A series of L-Seryl Aminoalkyl Glucosaminide Phosphate compounds (AGPs)was prepared as described in U.S. Pat. No. 6,113,918, issued Sep. 5,2000, and in U.S. patent application Ser. No. 09/439,839; filed Nov. 12,1999, now U.S. Pat. No. 6,303,347, each of which is incorporated hereinby reference in its entirety.

A dose of 20 μg of L-Seryl AGPs (RC-526, RC-554, RC-555, RC-537, RC-527,RC-538, RC-560, RC-512 and vehicle only) was given to groups of femaleBALB/c mice by intranasal (i.n.) administration. During the initial 4days following AGP administration, the mice were monitored for threesubjective indicators of disease (i.e. disease index) includingobserving ruffled fur, hunched posture and labored breathing. Theresults of these experiments is presented in FIG. 2. These dataindicated that i.n. administration of the RC-537, RC-527, RC-538 andRC-560 induce some toxicity in mice at the given dose of 20 μg.

Example 4 Clinical Symptoms Following Intranasal Administration ofL-Seryl Aminalkyl Glucosaminide Phosphates (AGPS) and InfluenzaChallenge

A dose of 20 μg L-Seryl AGPs (RC-526, RC-554, RC-555, RC-537, RC-527,RC-538, RC-560, RC-512 and vehicle only) was given to groups of femaleBALB/c mice by intranasal (i.n.) administration 2 days prior to or theday of lethal influenza challenge. All mice were challenged withapproximately 2 LD50 infectious influenza A/HK/68 administered i.n. Thedisease index (ruffled fur, hunched posture and labored breathing) wasmonitored during days 4-19 following influenza challenge. Weight lossand mortality were monitored for 21 days following influenza challenge.The results of these experiments are presented in FIG. 3.

These data demonstrated the efficacy of AGP compounds RC-538, RC560 andRC-512 in providing substantial protection against influenza challenge.

L-Seryl AGPs having 14 carbon fatty acid chains in the primary fattyacid position and 6 to 14 carbon fatty acids chains in the secondaryfatty acid position, (RC-526, RC-554, RC-555, RC-537, RC-527, RC-538,RC-560, RC-512, see FIG. 10), or combinations of 6 or 10 carbon fattyacids in the three secondary fatty acid positions, (RC-570, RC-568,RC-567, RC-566, RC-565, RC-569, see FIG. 10) were tested against MPL inthe influenza challenge model as described above.

The results indicate that chain length and position have an effect onsurvival and disease severity. Treatment with AGP compounds havingsecondary fatty acid chains containing 9 or more carbons provided moreprotection against the influenza challenge than did those with smallerfatty acid chains. (FIG. 11). Length of the fatty acid chain rather thanthe dose administered, is most influential on survival, (FIG. 12).RC-526 (6 carbon chain) showed no difference in protection up to dosesof 40 μg. RC-527 (10 carbon chain) provided maximum protection over adose range of 2.5 μg to 40 μg. Additionally, those compounds having atleast two 10 carbon fatty acid chains (RC-565 and RC-569) were moreprotective than those having fewer. (FIG. 13).

Example 5 Comparison of RC552 and MPL Using Human Whole Blood Culturesand Mouse Splenic Cultures

This Example discloses cytokine induction by the synthetic lipid Acompound RC552 as compared to the modified natural substancemonophosphoryl lipid A (MPL) using human whole blood cultures and mousesplenocyte culture.

Lipid A compounds were tested by reconstitution in 0.2% triethanolaminein sterile water for irrigation, incubated at 56° C. and sonicated for2×10 minutes at 37° C. LPS O55B5 (Sigma-Aldrich; St Louis, Mo.) wasdiluted into PBS.

Compounds were added to 450 μl of human whole blood and incubated withagitation for 5 to 24 hours. Three donors were selected (FIGS. 4-6,donors A-C, respectively). Supernatants were collected by centrifugationand diluted to ½ with an equal volume of PBS. (This dilution was notconsidered a dilution factor for cytokine calculations). Cytokineelaboration was measured by ELISA (R&D Systems; Minneapolis, Minn.)using the required volume of supernatant at full strength or diluted asmuch as ten fold.

BALB/c, DBA/2 and C3H/HEJ mice were purchased from The JacksonLaboratory (Bar Harbor, Me.). Spleens were taken from the mice between2PM and 3 PM, and separate single cell suspensions were obtained foreach mouse strain. Red blood cells were lysed using Tris-ammoniumchloride solution (Sigma-Aldrich), cells were washed and counted usingTrypan Blue (Sigma-Aldrich) exclusion. One million splenocytes werecultured per well in 1.0 mL of culture medium. Splenic culture medium(SCM) was designed for 5 day or longer cultures of mouse splenocytes andconsisted of RPMI 1640 (Sigma-Aldrich) supplemented to 5% with fetalbovine serum (HyClone; Logan, Utah), 100 ug/mL Gentamicin(Sigma-Aldrich), 250 ng/mL amphothericin (InVitrogen Life Technologies;Carlsbad, Calif.), 1×ITS (bovine insulin 500 ng/mL, human transferring500 ng/mLg, sodium selenite 250 ng/mL, Sigma), beta-mercaptoethanol 43nM (Sigma-Aldrich) purivic acid 1 mM (Sigma-Aldrich), HEPES 10 mM(Sigma-Aldrich). Data from these experiments is presented herein as FIG.8.

Using human whole blood cultures, four cytokines were measured: IL-10,MIP-1 beta, TNF alpha, and IL-8. Two donors were tested once and onedonor was tested twice. It is noteworthy that the donor who was testedtwice had very high background TNF alpha at one test, but very lowbackground TNF alpha one month later. Significant, however, is the timespan of culture. High background TNF alpha was obtained with a 5.5 hourculture, and low background TNF alpha was obtained with an overnight,about 24 hour, culture. Nonetheless, even the 5.5 hour-low backgroundculture was higher (about 600 pg/ml) than obtained in some previouscultures (518.2 pg/ml in a 5 hr test; 417 pg/ml in a 4 hr test; zeropg/ml 4 hr, low responder).

RC552 was similar to MPL for elaboration of TNF alpha in two of three 24hours cultures, and one 5.5 hour culture. IL-8 induction, however, byRC552 was different than that for MPL in three of three cases. IL-8induction by RC552 was lessened for two overnight cultures compared toMPL, but greater than MPL in one overnight culture.

For overnight cultures, IL-10 induction by RC552 was less than that forMPL. MIP-1 beta induction was lessened in one of 3 cases of overnightculture.

BALB/c responses and C3H/HEJ responses were compared for MPL and RC552.C3H/HEJ mice are genetic hyporesponders to LPS due to a mutation intoll-receptor 4. In these cultures, an oligonucleotide stimulant wasused as a positive control for C3H/HEJ cultures. This oligonucleotideinduced large amounts of IL-6 in BALB/c mice (1000 pg/mL) and in C3H/HEJmice (488.5 pg/mL). Similarly, MIP-1 beta was induced in BALB/c andC3H/HEJ cultures (589 pg/mL and 554 pg/mL), as was IL-10 (342 pg/mL and609 pg/mL), and TNF alpha (204 pg/mL and 30 pg/mL) in response to 10ug/mL MPL or RC552, respectively.

Neither MPL nor RC552 induced a cytokine response using C3H/HEJsplenocytes. In BALB/c splenocyte cultures, however, IL-10, MIP-1 beta,TNF alpha and IL-6 were induced. RC552 induced less MIP-1 beta, TNFalpha and IL-6 than did MPL at the same concentrations. RC552 inducedvery little IL-10 (10.4 to 11.6 pg/mL) compared to MPL (1144.1 to 176.6pg/mL).

When tested in a solution of 0.2% triethanolamine, RC552 has a similarbut not identical pro-inflammatory profile for TNF alpha induction asdoes MPL in two of three overnight cultures and one short-term cultureof human whole blood. See, FIGS. 4-6 (overnight cultures for donors A-C,respectively) and FIG. 7 (short-term culture for donor A). In addition,MIP-1 beta induction by RC552 was similar in two of three overnightcultures. Lessened IL-10 was induced by RC552 than MPL in one overnightculture. IL-8 induction was different than that for MPL in all casestested.

Using receptor deficient mice, it was clear that RC552 signals viatoll-like receptor 4. Using BALB/c mice that are lipid A responsive,RC552 induced a lessened cytokine profile at the concentrations tested.Interestingly, the concentrations tested were at the high end of a doseresponse relationship, and RC552 induced slightly greater MIP-1 beta andTNF alpha at the lower concentration (10 μg/mL) than at the higherconcentration (20 μg/mL) tested.

By comparing the human and mouse cytokine profiles, synthetic lipid Acompound RC552 lessened capacity for IL-10 induction in 2 day mousesplenocyte cultures and in 1 of 3 human blood cultures overnight, whentested at high concentrations of stimulant. In general, less TNF alphawas induced in overnight human blood cultures by RC552 than MPL. Aboutequal TNF alpha levels were induced in short term (5.5 hour) cultures ofhuman blood by RC552 compared to MPL. Microarray data using RNA obtainedfrom human macrophage stimulated with RC552 and MPL indicated early (1hour) TNF alpha RNA for both compounds, and no late TNF alpha RNA forboth compounds. RC552, however, induced very little 6 hour TNF alpha asopposed to MPL which had measurable 6 hour RNA.

In a separate set of experiments anticoagulated human whole blood wasincubated in the presence of various AGPs and the supernatants weretested for quantities of cytokines. Into sterile 1.4 mL microtubes in a96 well format (Matrix) were delivered 480 μL of human whole blood, andup to 20 μl of each AGP. Those AGPs tested included, L-Seryl AGPcompounds varying in secondary fatty acid chain length (RC-526, RC-554,RC-555, RC-537, RC-527, RC-538, RC-560, RC-512) and L-Seryl AGPs havingvarious combinations of 6 and 10 chain fatty acids (RC-570, RC-568,RC-567, RC-566, RC-565, RC-569, aminoethyl AGPs (RC-523, RC-524, RC-529,RC-577, see FIG. 10), L-serinamide AGPS(RC-522, and RC-515, see FIG.10), and Serinol AGPs, (RC-545, RC-574, RC-519, RC-541, RC-540, andRC-517 see FIG. 10), and AGPs varying in backbone length, RC-529 (2carbon linker), RC-525 (3 carbon linker), RC-557 (4 carbon linker),RC-571 (6 carbon linker).

The tubes were sealed with caps supplied by the manufacturer, and the 96well plate apparatus was placed on its side so that the microtubes werehorizontal, placed upon a “belly dancer” agitation platform, andcultured at 37° C. overnight, for 22 to 28 hours. At the conclusion ofculture, the tubes with holder were centrifuged momentarily to removeblood from the caps of each tube, the tubes were opened and 500 μl ofPBS were added, tubes sealed again, and inverted to mix. The addition of500 μL of PBS facilitated centrifugation as well as recovery of plasmasupernatants. Tubes in a plate holder were then centrifuged for 10minutes at 1800 RPM in an IEC Centra 8R centrifuge. Two aliquots of 310μL were then removed from each tube and stored frozen in a 96 wellformat until assayed for cytokine content. Cytokines (IFN, IL-2, IL-4,IL-5, IL-6, IL-8, MIP-1β, TNFα) were evaluated by ELISA (R and DSystems) and by cytometric bead array (BD Biosciences).

For each group of AGPs tested, two blood donors were used. Donor 1 andDonor 2 were not the same in all experiments reported in Tables 3 and 4.Each horizontal grouping represents one experiment in which donor 1 anddonor 2 were the same for all tests within that group. As a positivecontrol, separate samples of the same donor's blood were stimulated withpurified phytohaemagluttinin (PHA, Sigma), LPS from E coli O55B5(Sigma), E. coli DNA (Sigma) as well as MPL at doses up to 2011.Controls were used to determine the background state of the blood cellsas well as the ability of the blood cells to be stimulated.

In general, results compare favorably with those found in influenzachallenge model and the Listeria protection model, described herein.AGPs that were not active in these models did not induce cytokines inhuman blood cultures. AGPs that were weak protectors in mice, were weakinducers of cytokines in human whole blood cultures. AGPs that were verypotent in mouse protection studies also were potent inducers of humancytokines. Table 3 summarized the results for cytokines IL-6, IL-8,MIP-1β, TNFα, IL-10 and IFNγ. The maximum level of cytokine obtained isreported. TNFα and IFNγ are induced early have peaked before the assayis performed at 24 hours. In addition, IFNγ has not been found in humanblood cultures in a routine fashion, only sporadically. IFNγ inductionwas found in an isolated case of IFNγ expression. Background IFNγ levelsfor that donor were high (>200 pg/mL), suggesting that an immuneresponse was in process.

Table 4 provides the maximum levels of IL-4, IL-2 and IL-5 obtained, aswell as ratios of TNF to IL-10, and ratios of IL-10 to TNF. Inhibitiondata for IL-8, reported as a percentage, are also provided on the righthand side of Table 4. For example, 90% inhibition would calculate to a90% reduction of cytokine levels. MIP-10 results were similar.

RC-526, RC-554, RC-555, RC-570, RC-568, RC-567, and RC-566 exhibitedlittle if any cytokine stimulation but when added to Ogawa P. gingivalisLPS stimulated whole blood cells they inhibited LPS production ofcytokines 90-100%. See Table 4. Such LPS blocking would be useful forapplications where reduced LPS levels are desired. Topical applicationof such compounds would be useful to reduce bacteria levels prior totreatment, such as for dental surgery.

The results for RC-529 (2 carbon linker), RC-525 (3 carbon linker),RC-557 (4 carbon linker), RC-571 (6 carbon linker) suggest that as theaglycone carbon chain length increased, the cytokine stimulatory effectdecreased, indicating that greatest stimulation was achieved when thefatty acid residues attached to the aglycone moiety were closelyadjacent to the fatty acids attached to the glucosamine moiety.

TABLE 3 AGP IL-6 IL-8 MIP-1β TNFα cba IL-10 IFNγ cba RC# donor 1 donor 2donor 1 donor 2 donor 1 donor 2 donor 1 donor 2 donor 1 donor 2 donor 1donor 2 570 − − − − − − + − − 568 − − − − − +/− + − − 567 − − − − −+/− + − − 566 − − − − + + + − − 565 − + ++ ++ ++ +++ +/− − 569 + +/− +++++ ++ +++ ++++ + − 539 ++ + +++ ++++ +++ +++ ++++ +++ − 562 +/− − + +++++ ++ ++++ + − MPL + +/− +++ ++++ +++ +++ ++++ + − None − − − − − − − −− LPS + + ++ +++ +++ ++++ ++++ +++ − EcDNA + +/− ++ − +++ + ++++ − −PHA + + ++ +++ +++ ++++ ++++ ++++ + 526 − − − − − − − − ++ 554 − − − −+/− − − − ++ 555 − + +/− +/− +/− − − − ++ 537 + ++ ++ ++++ ++++ + ++ −++ 527 + ++ + ++++ ++++ +++ +++ − ++ 538 + ++ + ++++ ++++ +++ +++ − ++560 + ++ + ++++ ++++ ++ +++ − ++ 512 + ++ + ++++ +++ − ++ − ++ MPL +++ + +++ ++ − + − + None − − − − − − − − − LPS + +++ + +++ ++ ++ +++ −++ EcDNA + +++ + +++ +++ +++ +++ − + PHA +/− +/− +/− − +/− ++ + +++ ++523 + − +++ − + + − − − − +/− − 524 ++ + ++++ ++ + +++ − − + + +/− − 529++ + ++++ ++ + ++ − − + − +/− − 525 ++ + ++++ +++ ++ ++++ + + + + +/− −557 ++ + ++++ + ++ ++ − − + − − − 571 + − +++ − + + − − − − − − 577 ++/− +++ + + + − − − − − − MPL + + +++ + + ++ − − − − − − None − − − − −− − − − − − − LPS ++ + +++ + +++ ++++ + + + ++ +/− + EcDNA ++ ++ + +++++ ++++ − − ++ ++ − − PHA + + +++ ++ + ++ − − − − +/− ++ 522 +++ ++++++ +++ +++ +++ + + − − 515 +++ +++ ++++ ++++ +++ ++ + − − − 545 ++ +++++ +++ ++ +++ − − − − 544 +++ + ++++ +++ +++ ++ ++ − − − 519 ++ + +++++++ ++ ++ − + − − 541 +++ ++ ++++ ++ ++ ++ − − − − 540 ++ ++ ++++ ++++++ ++ + +/− − − 517 +++ +++ ++++ ++++ ++ ++ +/− +/− − − MPL + ++ ++++++ + ++ − − − − None − − − − − − − − − − LPS ++ +++ ++++ ++++ ++++ +++++++ +/− − − EcDNA ++ +++ ++++ ++++ +++ ++ ++++ + − − PHA +++ +++ ++++++++ ++++ +++ +++ − +++ ++

TABLE 4 10, 5, 2.5 ug/mL AGP TNFα/IL10 cba IL10/TNFα cba % inhib. IL-1βcba2 RC# IL-4 IL-2 IL-5 donor 1 donor 2 donor 1 donor 2 LPS IL-8 donor 1donor 2 570 − − − 100 568 − − − 96 567 − − − 100 566 − − − 93 565 − − −16 569 − − − −13 539 − − − 562 − − − MPL − − − None − − − LPS − − −EcDNA − + − PHA − − − 526 − − − 0.9 1.2 99 256 554 − − − 0.9 1.1 100 545555 − − − 0.8 1.2 99 519 537 − − − 0.8 9.7 1.3 0.3 33 8353 527 − − − 2.53.4 0.4 0.3 5 7102 538 − − − 2.3 4 0.5 0.3 −3 7767 560 − − − 0.8 5.1 1.30.2 −1 5833 512 − − − 3.9 0.4 −1 3230 MPL − − − 1 1 1213 None − − − 0LPS − − − 0.6 6.3 1.7 0.2 4414 EcDNA − − − 8.7 0.1 9453 PHA − ++ − 0.91.1 2023 523 − − − −47 524 − − − 0.2 0.3 1.8 4 − 529 − − − 0.4 6.2 − 525− − − 1.8 1.2 3.7 0.7 − 557 − − − 0.6 0.5 0.6 − 571 − − − 0.5 8.6 − 577− − − − MPL − − − None − − − 0.3 LPS − − − 0.1 3.2 2.5 EcDNA − − − 0.48.3 3.3 PHA − ++ − 522 − − − 0.4 2.7 515 − − − 0.4 0.8 3 1.4 545 − − −544 − − − 0.6 1.7 519 − − − 541 − − − 540 − − − 0.2 0.2 5.5 5.2 517 − −− 0.3 0.7 3.9 1.4 MPL − − − None − − − LPS − − − 1.7 0.3 0.7 4 EcDNA − −− 1.8 0.5 0.6 2.7 PHA − ++ −

Example 6 RC529 Stimulatory Capabilities Compared to MPL and RC552

This Example demonstrates that RC529 has superior immune stimulatorycapabilities as compared to MPL when assessed by IL-6, IL-10 and MIP-1beta elaboration from human peripheral blood mononuclear cells (PBMC).In contrast, IL-8 elaboration was similar to that of MPL.

PBMC were stored frozen until used. PBMC donor designation was AD112.PBMC at a density of 6.26×10⁵ were plated per well in a 48 well plate in1.0 ml of medium. Medium consisted of RPMI-1640 plus sodium bicarbonate,10% fetal bovine serum, 4 mM glutamine, 100 ug/ml gentamicin and 10 mMHEPES. PBMC were cultured for 22 hours at 37° C. in a carbon dioxideincubator. Supernatants were harvested and tested by ELISA (R&D Systems)for IL-6, IL-8, IL-10 and MIP-1 beta concentration. Cytokineconcentration in supernatants was compared to supernatants obtained fromunstimulated PBMC cultured identically.

At the doses tested, RC529 did not achieve dose-responsiveness at thelowest dose for IL-6 or IL-8. Compared to MPL, RC529 induced more IL-6,IL-10 and MIP-1 beta than did MPL. A disaccharide compound, RC552 wasgenerally intermediate in stimulatory capability on a mass basis. See,FIG. 9. These data show that RC529 is a strong inducer of IL-6, IL-10and MIP-1 beta from frozen human PBMC.

The same group of L-Seryl AGP compounds (RC-526, RC-554, RC-555, RC-537,RC-527, RC-538, RC-560, RC-512) and combined 6 and 10 chain fatty acids(RC-570, RC-568, RC-567, RC-566, RC-565, RC-569) as described above weretested and gave a result similar to that described using whole bloodculture assay as described in Example 5. Those AGP compounds havingsecondary fatty acid chains of 9 carbons or more, or at least two 10carbon fatty acids preferentially stimulated IL-6, IL-8, IL-10, andMIP-1β.

Example 7 Cell Surface Activation Markers

This experiment describes lineage specific cell surface activationmarkers in human PBMCs activated with LPS or selected AGPs. PBMCs wereharvested from a normal donor and plated at 1×10⁷ cells/well in a 16wells (6 well plate) with 3 ml RPMI. LPS (10 ng/ml), RC-526 (10 μg/ml)or RC-527 (10 μg/ml) were added to each of 4 wells. Cells were harvestedfrom each of 2 wells at 4 hr and 24 hr following activation andimmediately stained for the lineage and cell surface activation markers.

Table 5 shows the percent expression of cell surface activation markersin the indicated cell lineage. Table 6 shows the mean fluorescentintensities of the indicated activation marker within each cell subset.This table only includes the cell surface activation markers that showedsignificant changes in the expression following a 4 hr or 24 hrincubation with LPS, RC-527 or RC-526.

TABLE 5 No Activation LPS RC-527 RC526 4 hr activation T-cells (CD3+)CD69+ 1 5 10 1 Mono/Macs (CD14+) CD69+ 2 19 22 1 CD25+ 1 1 1 1 TLR2+ 5167 85 93 B-cells (CD19+) CD69+ 4 39 56 11 CD25+ 0 0 0 0 CD86+ 3 4 8 4CD95+ 6 6 7 6 NK-Cells (CD56+) CD69+ 4 34 40 4 CD3+ CD69+ 2 11 11 1 24hr activation T-Cells (CD3+) CD69+ 3 8 10 2 Mono/Macs (CD14+) CD14+CD69+ 5 18 22 2 CD14+ CD25+ 1 90 70 1 CD14+ TLR2+ 18 80 97 79 B-Cells(CD19+) CD19+ CD69+ 15 49 66 22 CD19+ CD25+ 5 35 34 9 CD19+ CD86+ 17 4051 22 CD19+ CD95+ 14 71 73 15 NK-Cells (CD56+) CD69+ 4 42 50 4 CD3+CD69+ 2 11 16 2

TABLE 6 MFI % No RC- Expression Activation LPS 527 RC526 4 hr activationMono/Macs (CD14+) CD11b+ 100% 2283 1650 1870 2154 CD54+(ICAM-1) 100% 559876 743 502 B-cells (CD19+) CD54+ (ICAM-1) 100% 57 57 65 55 24 hractivation Mono/Macs (CD14+) CD11b+ 100% 1737 893 1611 1986 CD54+(ICAM-1) 100% 1452 5290 4445 1590 B-cells (CD19+) CD54+ (ICAM-1 100% 110205 211 118 Note: Cells from the 4 hr and 24 hr activation groups werestained and acquired on different days so the comparisons of MFI betweentime points is not suggested.

Example 9 Murine Listeria monocytogenes Challenge Model

This example provides experiments evaluating the induction ofnon-specific resistance in the murine Listeria monocytogenes challengemodel performed using various AGPs and MPL. Mice (5 per group) weretreated intravenously with the 1 μg of an AGP or MPL solublized in 0.2%TEOA. Two days later the mice were challenged intravenously with a ˜10⁵L. monocytogenes 10403 serotype (provided by M. L. Gray, Montana StateUniversity, Bozeman, Mont.). Two days after the challenge, the mice weresacrificed and the number of colony forming units (CFUs) in the spleensof individual mice were determined by plating 10-fold serial dilutionsof splenic homogenates on tryptic soy agar plates. The degree ofprotection afforded by a given AGP or MPL was calculated by subtractingthe average number of bacteria per spleen (log10 value) in the group ofmice treated with a given compound, from the average number of bacteriaper spleen (log10 value) in a control group that was “sham” treated withvehicle (0.2% TEOA) prior to challenge with L. monocytogenes.

The same group of L-Seryl AGP compounds (RC-526, RC-554, RC-555, RC-537,RC-527, RC-538, RC-560, RC-512) and combined 6 and 10 chain fatty acids(RC-570, RC-568, RC-567, RC-566, RC-565, RC-569), as described above,were tested. As was seen in the influenza model, those AGPs having fattyacids of 9 or more carbons in the secondary position provided thegreatest protection, see FIG. 14. Those AGPs having at least two 10carbon fatty acid chains in the secondary position were only slightlyless protective than RC-527 which has three 10 carbon fatty acid chains,and were more protective than MPL, see FIG. 15.

AGPs having 10-carbon fatty acids in the secondary position from variousfamilies were also tested. L-Seryl (RC-527), Pyrrolidinomethyl (RC-590),Aminoethyl (RC-524), Serinamides (RC-522), Serinols and Serinolregioisomers (RC-540, RC-541, and RC-545) and miscellaneous other(RC-547, RC-558 and RC-573) AGPs provided protection that was equal toor greater than that provided by MPL, see FIG. 16.

AGPs varying in linker length RC-529(2 carbon linker), RC-525(3 carbonlinker), RC-557 (4 carbon linker), and RC-571 (6 carbon linker) againshowed that the greatest protection was achieved when the fatty acidresidues attached to the aglycone moiety were closely adjacent to thefatty acid residues attached to the glucosamine moiety, see FIG. 17.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference. Although the foregoing invention has beendescribed in some detail by way of illustration and example for purposesof clarity of understanding, it will be readily apparent to those ofordinary skill in the art in light of the teachings of this inventionthat certain changes and modifications may be made thereto withoutdeparting from the spirit or scope of the appended claims.

1. A method for ameliorating an infectious disease, autoimmune diseaseor allergic condition in a subject in need thereof comprising contactingthe subject with an effective amount of one or more compounds having theformula:

and pharmaceutically acceptable salts thereof, wherein X is a memberselected from the group consisting of —O— and —NH—; R¹ and R² are eachmembers independently selected from the group consisting of(C₂-C₂₄)acyl; R³ is a member selected from the group consisting of —Hand —PO₃ R¹¹ R¹², wherein R¹¹ and R¹² are each members independentlyselected from the group consisting of —H and (C₁-C₄)alkyl; R⁴ is amember selected from the group consisting of —H, —CH₃ and —PO₃ R¹³ R¹⁴,wherein R¹³ and R¹⁴ are each members independently selected from thegroup consisting of —H and (C₁-C₄)alkyl; and Y is a radical having theformula:

wherein the subscripts n, m, p and q are each independently an integerof from 0 to 6; R⁵ is (C₂-C₂₄)acyl; R⁶ and R⁷ are members independentlyselected from the group consisting of H and CH₃; R⁸ and R⁹ are membersindependently selected from the group consisting of H, OH,(C₁-C₄)alkoxy, —PO₃ H₂, —OPO₃ H₂, —SO₃ H, —OSO₃ H, —NR¹⁵ R¹⁶,—SR¹⁵, —CN,—NO₂,—CHO, —CO₂ R¹⁵, —CONR¹⁵ R¹⁶, —PO₃ R¹⁵ R¹⁶, —OPO₃ R¹⁵ R¹⁶, —SO₃ R¹⁵and —OSO₃ R¹⁵ wherein R¹⁵ and R¹⁶ are each members independentlyselected from the group consisting of H and (C₁-C₄)alkyl; and Z is —O—or —S—; with the proviso that when R³ is —PO₃ R¹¹ R¹², R⁴ is other than—PO₃ R¹³ R¹⁴, wherein the one or more compounds is administered in theabsence of exogenous antigen.
 2. A method in accordance with claim 1 forameliorating an infectious disease.
 3. A method in accordance with claim2 wherein the infectious disease is caused by a bacteria selected fromthe group consisting of Pseudomonas, Escherichia, Klebsiella,Enterobacter, Proteus, Serratia, Candida and Staphylococcus.
 4. A methodin accordance with claim 2 wherein the infectious disease is pneumonia.5. A method in accordance with claim 4, wherein said pneumonia isnosocomial pneumonia.
 6. A method in accordance with claim 4, whereinsaid pneumonia is in an HIV-positive patient.
 7. A method in accordancewith claim 2 wherein said infectious disease is a chronic infection. 8.A method in accordance with claim 5, wherein said chronic infectioncomprises chronic hepatitis, human papillomavirus, oral or vaginalcandidiasis, periodontal disease or chronic rhinosinusitis due to fungalcolonization.
 9. A method in accordance with claim 1 for ameliorating anautoimmune disease.
 10. A method in accordance with claim 9, whereinsaid autoimmune disease is selected from the group consisting ofinflammatory bowel disease, rheumatoid arthritis, chronic arthritis,multiple sclerosis and psoriasis.
 11. A method in accordance with claim10, wherein the autoimmune disease is inflammatory bowel disease.
 12. Amethod in accordance with claim 1, wherein two of said R¹, R² and R⁵ areselected from the group consisting of (C₂-C₆)acyl and the total numberof carbon atoms in R¹, R² and R⁵ is from about 6 to about
 22. 13. Amethod in accordance with claim 12, wherein the total number of carbonatoms in R¹, R² and R⁵ is from about 12 to about
 18. 14. A method inaccordance with claim 9, wherein two of said R¹, R² and R⁵ areindependently selected from the group consisting of (C₂-C₆)acyl and thetotal number of carbon atoms in R¹, R² and R⁵ is from about 6 to about22.
 15. A method in accordance with claim 14, wherein the total numberof carbon atoms in R¹, R² and R⁵ is from about 12 to about
 18. 16. Amethod in accordance with claim 1 for ameliorating an allergiccondition.
 17. A method in accordance with claim 16, wherein saidallergic condition is selected from the group consisting of asthma,atopic dermatitis, seasonal allergic disorder and chronicrhinosinusitis.
 18. A method in accordance with claim 16, wherein R¹, R²and R⁵ are independently selected from (C₇-C₁₁)acyl.
 19. A method inaccordance with claim 1, wherein said compound is administered to saidsubject by a route selected from the group consisting of parenteral,oral, intravenous, infusion, intranasal, inhalation, transdermal andtransmucosal administration.
 20. A method in accordance with claim 1,wherein at least two of said R¹, R² and R⁵ are selected from the groupconsisting of (C₂-C₆)acyl.
 21. A method in accordance with claim 1,wherein two of said R¹, R² and R⁵ are independently selected from thegroup consisting of (C₂-C₆)acyl and the total number of carbon atoms inR¹, R² and R⁵ is from about 6 to about
 22. 22. A method in accordancewith claim 1, wherein two of said R¹, R² and R⁵ are independentlyselected from the group consisting of (C₂-C₆)acyl and the total numberof carbon atoms in R¹, R² and R⁵ is from about 12 to about
 18. 23. Amethod in accordance with claim 1, wherein X and Z are both —O—.
 24. Amethod in accordance with claim 1, wherein R¹, R² and R⁵ are eachindependently selected from the group consisting of (C₁₂-C₂₄)acyl withthe proviso that the total number of carbon atoms in R¹, R² and R⁵ isfrom about 44 to about
 60. 25. A method in accordance with claim 24,wherein said total number of carbon atoms is from about 46 to about 52.26. A method in accordance with claim 24, wherein X and Z are both —O—.27. A method in accordance with claim 1, wherein R¹, R² and R⁵ areindependently selected from (C₇-C₁₁)acyl.
 28. A method in accordancewith claim 1, wherein the compound is in the form of a pharmaceuticallyacceptable salt.
 29. A method for the therapeutic treatment of abacterial or viral infection in a subject comprising contacting thesubject in need thereof with an effective amount of one or morecompounds having the formula:

and pharmaceutically acceptable salts thereof, wherein X is a memberselected from the group consisting of —O— and —NH—; R¹ R² are eachmembers independently selected from the group consisting of(C₂-C₂₄)acyl; R³ is a member selected from the group consisting of —Hand —PO₃ R¹¹ R¹², wherein R¹¹ R¹² are each members independentlyselected from the group consisting of —H and (C₁-C₄)alkyl; R⁴ is amember selected from the group consisting of —H, —CH₃ and —PO₃ R¹³ R¹⁴,wherein R¹³ and R¹⁴ are each members independently selected from thegroup consisting of —H and (C₁-C₄)alkyl; and Y is a radical having theformula:

wherein the subscripts n, m, p and q are each independently an integerof from 0to 6; R⁵ is (C₂-C₂₄)acyl; R⁶ and R⁷ are members independentlyselected from the group consisting of H and CH₃; R⁸ and R⁹ are membersindependently selected from the group consisting of H, OH,(C₁-C₄)alkoxy, —PO₃ H₂, —OPO₃ H₂, —SO₃ H, —OSO₃ H, —NR¹⁵ R¹⁶, —SR¹⁵,—CN, —NO₂,—CHO, —CO₂ R¹⁵, —CONR¹⁵ R¹⁶, —PO₃ R¹⁵ R¹⁶, —OPO₃ R¹⁵ R¹⁶, —SO₃R¹⁵ and —OSO₃ R¹⁵ wherein R¹⁵ and R¹⁶ are each members independentlyselected from the group consisting of H and (C₁-C₄)alkyl; and Z is —O—or —S—; with the proviso that when R³ is —PO₃ R¹¹ R¹², R⁴ is other than—PO₃ R¹³ R¹⁴, wherein the one or more compounds is administered in theabsence of exogenous antigen.
 30. A method in accordance with claim 29,wherein said infection is a nosocomial infection.
 31. A method inaccordance with claim 30, wherein said nosocomial infection is apneumonia.
 32. A method in accordance with claim 29, wherein saidinfection is in an HIV-positive patient.
 33. A method in accordance withclaim 32, wherein the infection in said HIV-positive patient ispneumonia.
 34. A method in accordance with claim 33, wherein saidinfection is caused by P. carinii.
 35. A method in accordance with claim29, wherein the compound is in the form of a pharmaceutically acceptablesalt.
 36. A method in accordance with claim 1, wherein said subject isan immunocompromised subject.
 37. A method in accordance with claim 29,wherein said subject is an immunocompromised subject.
 38. A method inaccordance with claim 28, wherein said subject is an immunocompromisedsubject.
 39. A method in accordance with claim 1, wherein said subjectis one having chronic obstructive pulmonary disease.
 40. A method inaccordance with claim 29, wherein said subject is one having chronicobstructive pulmonary disease.
 41. A method in accordance with claim 28,wherein said subject is one having chronic obstructive pulmonarydisease.
 42. A method in accordance with claim 29, wherein saidinfection comprises a bacterial infection.
 43. A method in accordancewith claim 42, wherein said subject is an immunocompromised subject. 44.A method in accordance with claim 42, wherein said subject is one havingchronic obstructive pulmonary disease.
 45. A method according to claim28, wherein said infectious disease comprises a bacterial infection. 46.A method in accordance with claim 45, wherein said subject is animmunocompromised subject.
 47. A method in accordance with claim 45,wherein said subject is one having chronic obstructive pulmonarydisease.
 48. A method according to claim 29, wherein said infectioncomprises a viral infection.
 49. A method in accordance with claim 48,wherein said subject is an immunocompromised subject.
 50. A method inaccordance with claim 48, wherein said subject is one having chronicobstructive pulmonary disease.
 51. A method according to claim 28,wherein said infectious disease comprises a viral infection.
 52. Amethod in accordance with claim 51, wherein said subject is animmunocompromised subject.
 53. A method in accordance with claim 51,wherein said subject is one having chronic obstructive pulmonarydisease.